Purpose of methanol in the Blue Silver stain - (May/04/2009 )

May I know the purpose of methanol in the Blue Silver stain coz during the staining, a senior told me to stain the gels in a closed box (i.e. we put the staining tray in a zip lock bag), telling that it could prevent the methanol evaporation. But i read the paper and it did not mentioned the function of the methanol in the stain. Is there a need to do the staining this way?

Likewise, may i know whether u fix the gel before the Blue silver staining?

Thx very much.

-barnacleman-

Methanol fixes the proteins in the gel.

-bob1-

I use Blue Silver stain and gels seem to look a bit better if they're fixed before staining, however you get a bit more background.

-K.B.-

K.B. on May 9 2009, 08:13 AM said:

I use Blue Silver stain and gels seem to look a bit better if they're fixed before staining, however you get a bit more background.

Hi K.B., may i know the way u use to ensure all the blue silver particles are completely dissolved?

In our lab, we follow this sequence --> H3PO4 --> some H2O + Ammonium sulfate powder (stir overnight to ensure it is completely dissolved) --> CBB-G250 --> Methanol --> make up to 2L using water.

But for the CBB-G250, I added it on the next day morning, and stir it for ~ 1 - 2 hrs. However, some of the particles already stick quite hard to the bottle and can't dissolve. Is there a better way for this?

Likewise, I'd like to know if u will reuse the blue silver stain or prepare new ones each time.